Recombinant microorganism having ability to produce [lactate-co-glycolate] copolymer from glucose, and method for preparing [lactate-co-glycolate] copolymer using same

Abstract

There is provided a recombinant microorganism having producibility of poly(lactate-co-glycolate) from glucose, and more particularly, a recombinant microorganism having producibility of poly(lactate-co-glycolate) without adding an exogenous glycolate precursor, and a method of preparing [poly(preparing lactate-co-glycolate)] using the same. According to the present invention, the poly(lactate-co-glycolate) in which the concentration of the glycolate fraction is high may be prepared at a high concentration without supplying exogenous glyoxylate. Therefore, the present invention may be effectively used for treatment.

The invention claimed is: 
     
       1. A recombinant microorganism having producibility of poly(lactate-co-glycolate), wherein the microorganism is transfected so that a gene coding poly(hydroxyalkanoate) (PHA) synthase, a gene coding propionyl-CoA transferase, and a gene coding glycerate dehydrogenase (EC 1.1.1.26) are inserted. 
     
     
       2. The recombinant microorganism of  claim 1 , wherein the propionate CoA-transferase is propionate CoA-transferase of  C. propionicum  or Pct540, which is a mutant enzyme of propionate CoA-transferase of  C. propionicum.    
     
     
       3. The recombinant microorganism of  claim 1 , wherein the poly(hydroxyalkanoate) (PHA) synthase is PHA synthase of  Pseudomonas  sp. 6-19 or a mutant enzyme of PHA synthase of  Pseudomonas  sp. 6-19. 
     
     
       4. The recombinant microorganism of  claim 3 , wherein the gene coding the mutant enzyme of the PHA synthase has a base sequence corresponding to an amino acid sequence including at least one mutation selected from a group consisting of E130D, S325T, L412M, S477R, S477H, S477F, S477Y, S477G, Q481M, Q481K, and Q481R in an amino acid sequence of SEQ ID No: 1. 
     
     
       5. The recombinant microorganism of  claim 3 , wherein the gene coding the mutant enzyme of the PHA synthase has a base sequence corresponding to an amino acid sequence selected from a group consisting of:
 an amino acid sequence (C1335) in which E130D, S325T, L412M, S477G, and Q481M in the amino acid sequence of SEQ ID No: 1 are mutated; 
 an amino acid sequence (C1310) in which E130D, S477F, and Q481K in the amino acid sequence of SEQ ID No: 1 are mutated; and 
 an amino acid sequence (C1312) in which E130D, S477F, and Q481R in the amino acid sequence of SEQ ID No: 1 are mutated. 
 
     
     
       6. The recombinant microorganism of  claim 1 , wherein the gene coding glycerate dehydrogenase (EC 1.1.1.26) is derived from  Pasteurella multocida.    
     
     
       7. The recombinant microorganism of  claim 6 , wherein the gene coding glycerate dehydrogenase has a base sequence of SEQ ID No: 2. 
     
     
       8. A recombinant microorganism having producibility of poly(lactate-co-glycolate), wherein the microorganism is transfected so that a gene coding PHA synthase, a gene coding propionyl-CoA transferase, and a gene coding glycerate dehydrogenase (EC 1.1.1.26) are inserted, and a gene (iclR) coding isocitrate lyase regulator or aceB (malate synthase) gene is deleted. 
     
     
       9. The recombinant microorganism of  claim 8 , wherein the propionate CoA-transferase is propionate CoA-transferase of  C. propionicum  or Pct540, which is a mutant enzyme of propionate CoA-transferase of  C. propionicum.    
     
     
       10. The recombinant microorganism of  claim 8 , wherein the poly(hydroxyalkanoate) (PHA) synthase is PHA synthase of  Pseudomonas  sp. 6-19 or a mutant enzyme of PHA synthase of  Pseudomonas  sp. 6-19. 
     
     
       11. The recombinant microorganism of  claim 10 , wherein the gene coding the mutant enzyme of the PHA synthase has a base sequence corresponding to an amino acid sequence including at least one mutation selected from a group consisting of E130D, S325T, L412M, S477R, S477H, S477F, S477Y, S477G, Q481M, Q481K, and Q481R in an amino acid sequence of SEQ ID No: 1. 
     
     
       12. The recombinant microorganism of  claim 10 , wherein the gene coding the mutant enzyme of the PHA synthase has a base sequence corresponding to an amino acid sequence selected from a group consisting of:
 an amino acid sequence (C1335) in which E130D, S325T, L412M, S477G, and Q481M in the amino acid sequence of SEQ ID No: 1 are mutated; 
 an amino acid sequence (C1310) in which E130D, S477F, and Q481K in the amino acid sequence of SEQ ID No: 1 are mutated; and 
 an amino acid sequence (C1312) in which E130D, S477F, and Q481R in the amino acid sequence of SEQ ID No: 1 are mutated. 
 
     
     
       13. The recombinant microorganism of  claim 10 , wherein the gene coding glycerate dehydrogenase (EC 1.1.1.26) is derived from  Pasteurella multocida  or  E. coli.    
     
     
       14. The recombinant microorganism of  claim 13 , wherein the gene coding glycerate dehydrogenase has a base sequence of SEQ ID No: 2. 
     
     
       15. The recombinant microorganism of  claim 10 , wherein both of the gene (iclR) coding isocitrate lyase regulator and a gene (aceB) coding isocitrate lyase are deleted. 
     
     
       16. The recombinant microorganism of  claim 10 , wherein the gene (aceA) coding isocitrate lyase is additionally amplified. 
     
     
       17. A method of preparing poly(lactate-co-glycolate) (PLGA) including: culturing the recombinant microorganism of  claim 1  to produce poly(lactate-co-glycolate); and obtaining the produced PLGA. 
     
     
       18. A method of preparing poly(lactate-co-glycolate) (PLGA) including: culturing the recombinant microorganism of  claim 10  to produce poly(lactate-co-glycolate); and obtaining the produced PLGA.